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Team:KIT-Kyoto/Notebook/ATF1/may
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Revision as of 10:05, 26 September 2013
May 16th
We performed PCR to amplify the ATF1 gene.
Primers:
Forward:5’-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3’
Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’
Reaction composition is as follows:
|
Buffer |
50uL |
|
dNTP |
20uL |
|
Primer mix |
1uL |
|
DNA sample |
0.5uL |
|
KOD-FX |
2uL |
|
H2O |
26.5uL |
|
total |
100uL |
We could not get any PCR product.
May 17th -20th
Changed PCR conditions with another DNA template. We performed PCR of the ATF1 gene again.
Primers:
Forward
Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’
Reaction composition is as follows:
|
Buffer |
50uL |
|
dNTP |
20uL |
|
Primer mix |
1uL |
|
DNA sample |
0.5uL |
|
KOD-FX |
2uL |
|
H2O |
26.5uL |
|
total |
100uL |
We could not get any PCR product.
May 30th -31th
We transformed a plasmid carrying the ATF1 gene into E.coli.
This plasmid was obtained from iGEM DNA distributions kit 2012 (plate 2, o-7).
We could see approximately 50 colonies and cultured furthermore.
